动物造模,荧光定量PCR检测,诺如病毒 z-bio 2021-06-21 343

　　Objective: To establish a real-time fluorescent quantitative PCR detection method for murine norovirus (MNV), and to provide a basis for establishing a standardized detection method for MNV.

　　Method: Design specific primers based on the nucleic acid sequences of 60 MNV strains published by NCBI, construct standard positive quality control materials, establish MNV real-time fluorescent quantitative PCR method, and evaluate specificity, sensitivity, reproducibility and stability. This method is used to detect the cecum content samples of 766 mice submitted for inspection, and to get a preliminary understanding of the MNV infection in the Beijing experimental mice.

　　Result: We successfully established a real-time fluorescent quantitative PCR detection method for MNV. This method is highly specific and will not cross-react with the same species of human norovirus (HuNoV) and feline calicivirus (FCV). Detection sensitivity. Up to 101 copies/μL. The coefficient of variation of intra-assay and inter-assay CT values is less than 2? Using this method, 301 MNV nucleic acid-positive samples were detected in 766 mouse cecal contents samples, and the positive rate was 39.3?

　　Conclusion: The established real-time fluorescent quantitative PCR detection method for MNV has good specificity, high sensitivity and good reproducibility, and can be used for rapid quantitative detection of MNV.